1. Crystal Structure of Escherichia coli RNase D, an Exoribonuclease Involved in Structured RNA Processing 
Yuhong Zuo, Yong Wang, Arun Malhotra 
Structure 
Volume 13, Issue 7, Pages (July 2005)
DOI: /j.str
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2. Figure 1 Multiple Alignment of RNase D-Related Sequences
Sequences are from the NCBI nonredundant protein sequence database. Sequence included here are: RND_ECOLI, E. coli RNase D (accession no. G133152); RND_MYCTU, M. tuberculosis Rv2681 protein, a putative RNase D (G ); RND_AGRTU, putative A. tumefaciens RNase D (G ); RRP6_YEAST, S. cerevisiae RRP6 protein, an exosome component (G ); PMC2_HUMAN, the human RRP6 equivalent (G ); RND_RICPR, putative R. prowazekii RNase D (G ); RND_SYNY3, putative Synechocystis RNase D (G ); and RND2_AGRTU, another A. tumefaciens RNase D-related protein (G ). These sequences are chosen because they are representative: RRP6_YEAST and PMC2_HUMAN are eukaryotic RNase D homologs; RND_MYCTU and RND_AGRTU are two distant bacterial homologs of E. coli RNase D; the other three sequences are distantly related to E. coli RNase D, but lacking one (RND_RICPR) or both (RND_SYNY3 and RND2_AGRTU) C-terminal domains. This sequence alignment was initially generated by ClustalX and manually adjusted based on secondary structure predictions due to very weak sequence conservation, especially at the C-terminal domain. Many other related sequences (not shown) have been used to help produce this sequence alignment. The conserved DEDDy residues are marked with red triangles, and are distributed among three exo motifs (Zuo and Deutscher, 2001). Residues conserved among most RNase D homologs are boxed and colored. Highly conserved residues are highlighted in red. The numbering on the top is according to E. coli RNase D sequence.
Structure  , DOI: ( /j.str )
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3. Figure 2 Crystal Structure of E. coli RNase D
(A) A ribbon representation of RNase D in stereo. The three structural domains are colored differently: cyan = N-terminal DEDD domain; yellow = first HRDC domain (HRDC1); orange = C-terminal HRDC domain (HRDC2). Also shown are conserved DEDDy residues (red sticks) and bound metal ions (balls).
(B) Molecular surface of RNase D, colored by local electrostatic potential using GRASP (Nicholls et al., 1991). Red color indicates negative potential, blue color indicates positive potential. Also shown is the backbone of a double-stranded nucleic acid fragment manually modeled into this structure to display a possible mechanism of substrate binding.
(C) Crystal packing in RNase D crystal (stereo view). Shown here is a P unit cell of RNase D crystal. Molecule A in the middle is shown as a molecular surface color-coded as in (A); molecules B (lower left) and C (lower right) in a ribbon representation are colored purple; molecules D (upper front) and D′ (upper back) are separated by one-unit cell dimension, and are color-coded as in molecule A, but in a ribbon representation.
Structure  , DOI: ( /j.str )
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4. Figure 3 DEDD Exonuclease Domains
(A) Comparison of DEDD domains from various proteins. The E. coli RNase D structure is compared to the structures of Klenow fragment (PDB ID: 2KZZ), E. coli exonuclease 1 (EX1, PDB ID: 1FXX), N-terminal domain of DNA polymerase III ϵ subunit (DP3E, pDB ID: 1J53), and yeast POP2 protein (PDB ID: 1UOC). The DEDD core domains are colored purple in each ribbon diagram. RNase D and Klenow fragment belong to the DEDDy subgroup, EX1 and DP3E contain DEDDh motifs, whereas POP2 contains only one of the four highly conserved acidic residues. The views shown were obtained by superposing the full catalytic DEDD domains. The DEDDh/DEDDy or residues at equivalent positions are colored red and shown as sticks. An additional helix (colored cyan) shared between RNase D and Klenow fragment connects their DEDD domains to other structural domains.
(B) Superposition of the Cα traces of RNase D (in color) and Klenow fragment (in gray) DEDD domains is shown in stereo. Also shown are metal ions (balls) and DEDDy residues (red or gray sticks), as well as a dinucleotide (orange sticks) bound in the Klenow fragment crystals.
(C) Stereo view of the 2Fo − Fc density map (contoured at 1.0 σ) at the RNase D putative active center. Superimposed is a partial ball-and-stick model of RNase D. DEDDy residues and several water residues are labeled. “A” and “B” indicate two bound metal ions in RNase D crystal.
Structure  , DOI: ( /j.str )
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5. Figure 4
HRDC Domains
(A) Structure-guided sequence alignment of HRDC domains. HRDC1 = first HRDC domain in RND; HRDC2 = RND C-terminal HRDC domain; 1D8B = Sgs1p HRDC domain (PDB ID: 1D8B); 1GO3 = HRDC domain of an archaeal RNA polymerase (PDB ID: 1GO3). HRDC1 secondary structure is shown on top, HRDC2 secondary structure is shown on bottom, and every tenth residue is numbered according the E. coli RNase D sequence. The labels of secondary structures are the same as in Figure 1.
(B) Superposition in stereo of the Cα traces of HRDC1 (green), HRDC2 (cyan), and Sgs1p HRDC domain (purple).
(C) Molecular surface of HRDC1, HRDC2 and Sgs1p HRDC domain, colored on the same scale by local electrostatic potential using GRASP (Nicholls et al., 1991). The upper panel is shown in the same orientations as in (B). The lower panel shows a view rotated by 180° along the horizontal axis. Blue color represents positive potential; red color represents negative potential.
Structure  , DOI: ( /j.str )
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